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US 7,057,031 B2Granted
Nucleotide compositions comprising photocleavable markers and methods of preparation thereof
AssigneeAmberGen, Inc.
InventorsJerzy Olejnik, Edyta Krzymanska-Olejnik, Kenneth J. Rothschild
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Abstract
Labelled nucleotides and polynucleotides useful in the sequencing of nucleic acids are described. Methods of preparing photocleavable marker nucleotides and photocleavable marker-polynucleotide conjugates are described. Such photocleavable markere nucleotides can be incorporated into nucleic acid so as to create photocleavable marker-polynucleotide conjugates.
Claims11
A method of preparing a marker-photocleavable linker-nucleotide conjugate comprising:
- a) providing i) a photocleavable linker comprising a protective group, ii) an amino-nucleotide analog, and iii) an activated marker molecule selected from the group consisting of BODIPY-FL-SF and Cy5-NHS;
- b) operably linking said photocleavable linker to said amino-nucleotide analog to produce a photocleavable linker-nucleotide conjugate;
- c) removing said protective group from said photocleavable linker-nucleotide conjugate under conditions such that an activated photocleavable linker-nucleotide conjugate is created, wherein said activated photocleavable linker-nucleotide conjugate has an exposed reactive site on the linker portion of the conjugate;
- and
- d) contacting said activated marker molecule with said activated photocleavable linker-nucleotide conjugate under conditions such that a marker-photocleavable linker-nucleotide conjugate is produced.
2 more elements
A marker-photocleavable linker-nucleotide conjugate of the structure:
- wherein F is a fluorophore marker and PPP is:
Dependencies are inferred from the claim text.